Antibody Type: Recombinant Rabbit Monoclonal

Application: WB,IHC-P,IHC-Fr,IF,FCM

Reactivity: Human,Mouse,Rat

IgG

3P967

Human,Mouse,Rat

1. Paraformaldehyde-fixed, paraffin embedded (Human smooth muscle); Antigen retrieval by boiling in sodium citrate buffer (pH6.0) for 15 min; Block endogenous peroxidase by 3% hydrogen peroxide for 20 min; Blocking buffer (normal goat serum) at 37°C for 30 min; Antibody incubation with (YY1 (Nuclear Loading Control)) Monoclonal Antibody, Unconjugated (TMAB-01992) at 1:200 overnight at 4°C, followed by operating according to SP Kit (Rabbit) instructionsand DAB staining.
2. Paraformaldehyde-fixed, paraffin embedded (mouse lymphoid); Antigen retrieval by boiling in sodium citrate buffer (pH6.0) for 15 min; Block endogenous peroxidase by 3% hydrogen peroxide for 20 min; Blocking buffer (normal goat serum) at 37°C for 30 min; Antibody incubation with (YY1 (Nuclear Loading Control)) Monoclonal Antibody, Unconjugated (TMAB-01992) at 1:200 overnight at 4°C, followed by operating according to SP Kit (Rabbit) instructionsand DAB staining.
3. Paraformaldehyde-fixed, paraffin embedded (mouse ovary); Antigen retrieval by boiling in sodium citrate buffer (pH6.0) for 15 min; Block endogenous peroxidase by 3% hydrogen peroxide for 20 min; Blocking buffer (normal goat serum) at 37°C for 30 min; Antibody incubation with (YY1 (Nuclear Loading Control)) Monoclonal Antibody, Unconjugated (TMAB-01992) at 1:200 overnight at 4°C, followed by operating according to SP Kit (Rabbit) instructionsand DAB staining.
4. Paraformaldehyde-fixed, paraffin embedded (mouse brain); Antigen retrieval by boiling in sodium citrate buffer (pH6.0) for 15 min; Block endogenous peroxidase by 3% hydrogen peroxide for 20 min; Blocking buffer (normal goat serum) at 37°C for 30 min; Antibody incubation with (YY1 (Nuclear Loading Control)) Monoclonal Antibody, Unconjugated (TMAB-01992) at 1:200 overnight at 4°C, followed by operating according to SP Kit (Rabbit) instructionsand DAB staining.
5. Paraformaldehyde-fixed, paraffin embedded (mouse placenta); Antigen retrieval by boiling in sodium citrate buffer (pH6.0) for 15 min; Block endogenous peroxidase by 3% hydrogen peroxide for 20 min; Blocking buffer (normal goat serum) at 37°C for 30 min; Antibody incubation with (YY1 (Nuclear Loading Control)) Monoclonal Antibody, Unconjugated (TMAB-01992) at 1:200 overnight at 4°C, followed by operating according to SP Kit (Rabbit) instructionsand DAB staining.
6. Paraformaldehyde-fixed, paraffin embedded (Rat lymphoid); Antigen retrieval by boiling in sodium citrate buffer (pH6.0) for 15 min; Block endogenous peroxidase by 3% hydrogen peroxide for 20 min; Blocking buffer (normal goat serum) at 37°C for 30 min; Antibody incubation with (YY1 (Nuclear Loading Control)) Monoclonal Antibody, Unconjugated (TMAB-01992) at 1:200 overnight at 4°C, followed by operating according to SP Kit (Rabbit) instructionsand DAB staining.
7. Paraformaldehyde-fixed, paraffin embedded (Rat bladder); Antigen retrieval by boiling in sodium citrate buffer (pH6.0) for 15 min; Block endogenous peroxidase by 3% hydrogen peroxide for 20 min; Blocking buffer (normal goat serum) at 37°C for 30 min; Antibody incubation with (YY1 (Nuclear Loading Control)) Monoclonal Antibody, Unconjugated (TMAB-01992) at 1:200 overnight at 4°C, followed by operating according to SP Kit (Rabbit) instructionsand DAB staining.
8. Paraformaldehyde-fixed, paraffin embedded (Rat ovary); Antigen retrieval by boiling in sodium citrate buffer (pH6.0) for 15 min; Block endogenous peroxidase by 3% hydrogen peroxide for 20 min; Blocking buffer (normal goat serum) at 37°C for 30 min; Antibody incubation with (YY1 (Nuclear Loading Control)) Monoclonal Antibody, Unconjugated (TMAB-01992) at 1:200 overnight at 4°C, followed by operating according to SP Kit (Rabbit) instructionsand DAB staining.
9. Paraformaldehyde-fixed, paraffin embedded (rat brain); Antigen retrieval by boiling in sodium citrate buffer (pH6.0) for 15 min; Block endogenous peroxidase by 3% hydrogen peroxide for 20 min; Blocking buffer (normal goat serum) at 37°C for 30 min; Antibody incubation with (YY1 (Nuclear Loading Control)) Monoclonal Antibody, Unconjugated (TMAB-01992) at 1:200 overnight at 4°C, followed by operating according to SP Kit (Rabbit) instructionsand DAB staining.
10. Paraformaldehyde-fixed, paraffin embedded (rat breast); Antigen retrieval by boiling in sodium citrate buffer (pH6.0) for 15 min; Block endogenous peroxidase by 3% hydrogen peroxide for 20 min; Blocking buffer (normal goat serum) at 37°C for 30 min; Antibody incubation with (YY1 (Nuclear Loading Control)) Monoclonal Antibody, Unconjugated (TMAB-01992) at 1:200 overnight at 4°C, followed by operating according to SP Kit (Rabbit) instructionsand DAB staining.
11. Paraformaldehyde-fixed, paraffin embedded (rat placenta); Antigen retrieval by boiling in sodium citrate buffer (pH6.0) for 15 min; Block endogenous peroxidase by 3% hydrogen peroxide for 20 min; Blocking buffer (normal goat serum) at 37°C for 30 min; Antibody incubation with (YY1 (Nuclear Loading Control)) Monoclonal Antibody, Unconjugated (TMAB-01992) at 1:200 overnight at 4°C, followed by operating according to SP Kit (Rabbit) instructionsand DAB staining.
12. Paraformaldehyde-fixed, paraffin embedded (human breast carcinoma); Antigen retrieval by boiling in sodium citrate buffer (pH6.0) for 15 min; Block endogenous peroxidase by 3% hydrogen peroxide for 20 min; Blocking buffer (normal goat serum) at 37°C for 30 min; Antibody incubation with (YY1 (Nuclear Loading Control)) Monoclonal Antibody, Unconjugated (TMAB-01992) at 1:200 overnight at 4°C, followed by operating according to SP Kit (Rabbit) instructionsand DAB staining.
13. Blank control: K562. Primary Antibody (green line): Rabbit Anti-YY1 (Nuclear Loading Control) antibody (TMAB-01992)
Dilution: 1 μg/10^6 cells;
Isotype Control Antibody (orange line): Rabbit IgG.
Secondary Antibody: Goat anti-rabbit IgG-FITC
Dilution: 0.5 μg/test.
Protocol
The cells were fixed with 4% PFA (10 min at room temperature) and then permeabilized with 90% ice-cold methanol for 20 min at-20°C. The cells were then incubated in 5% BSA to block non-specific protein-protein interactions for 30 min at room temperature. Cells stained with Primary Antibody for 30 min at room temperature. The secondary antibody used for 40 min at room temperature.
14. Sample:
Lane 1: MCF-7 (Human) Cell Lysate at 30 μg
Lane 2: Jurkat (Human) Cell Lysate at 30 μg
Lane 3: Du145 (Human) Cell Lysate at 30 μg
Lane 4: U251 (Human) Cell Lysate at 30 μg
Lane 5: Panc-1 (Human) Cell Lysate at 30 μg
Lane 6: MDA-MB-231 (Human) Cell Lysate at 30 μg
Lane 7: Molt-4 (Human) Cell Lysate at 30 μg
Lane 8: Hela (Human) Cell Lysate at 30 μg
Lane 9: HL60 (Human) Cell Lysate at 30 μg
Lane 10: Urinary bladder (Mouse) Lysate at 40 μg
Lane 11: Testis (Mouse) Lysate at 40 μg
Primary: Anti-YY1 (TMAB-01992) at 1/1000 dilution
Secondary: IRDye800CW Goat Anti-Rabbit IgG at 1/20000 dilution
Predicted band size: 65-68 kDa
Observed band size: 65 kDa

WB,IHC-P,IHC-Fr,IF,FCM

WB: 1:500-2000; IHC-P: 1:50-200; IHC-Fr: 1:50-200; IF: 1:50-200; FCM: 1ug/Test

Monoclonal

Recombinant Protein: human YY1

Human

7528

Zinc Finger,Other factors,Transcription Factors,YY1,Neurogenesis